Tumorigenesis is associated with the disbalance of oncogenes and antioncogenes. For almost all of the oncogenes or antioncogenes, their functions and effects are finally converged to cell cycle. Therefore, tumor can be taken as a cell cycle disease (CCD), and it is one of the routes for treating tumor by regulating or blocking cell cycle. In current, it is found that there are a lot of molecules associated with cell cycle regulation, among which Cyclin-Dependent-Kinases (CDKs) are core molecules of cell cycle regulatory network. CDKs, as catalytic subunits, are a class of Ser/Thr kinases, which participate in different stages of cell cycle as important signaling molecules in cells. Studies show that in a cell cycle regulatory network with CDKs as core, any abnormity would result in abnormal cell cycle and finally result in tumorigenesis. CDK family now has 21 isoforms, which work by binding to their regulatory subunit cyclins. In addition to the role in regulating the cell cycle, CDK isoforms are also involved in regulating transcription, DNA repair, differentiation and programmed cell death. Based on the key role of CDKs in regulating proliferation and death of tumor cells, the family of CDKs provides a chance and a new field for the discovery and development of anti-tumor drugs.
In the development of drugs, the first generation of CDK inhibitors, represented by flavopiridol, UCN-01 and the like, are designated as “pan-CDK” inhibitors, which block all the isoforms of CDK family equivalently and exhibit a relatively high toxicity in clinical trial, and some of them cannot be administered in a therapeutically effective amount. Therefore, human begins to develop selective CDK inhibitors to enhance the selectivity of the therapy and prevent normal cells from injury by some side effects.
Among the CDK isoforms involved in cell cycle, CDK4/6 plays an unreplaceable role. Cancer-associated cell cycle mutations are mainly present in G1 phase and G1/S transition. The complex formed by CDK4/6 and Cyclin D releases the bound transcriptional factor E2F by phosphorylation (pRb) of the antioncogene product Rb, and triggers transcription of genes associated with S phase, thereby promoting cells to pass the checkpoint and to transit from G1 phase to S phase. About 80% of human tumors are abnormal in cyclin D-CDK4/6-INK4-Rb pathway. Due to the alteration of the pathway, the G1 phase is accelerated so that tumor cells have the proliferation sped up and thus acquire survival advantage. Therefore, the interference of the pathway has become a strategy of treatment, and CDK4/6 has become a new anti-tumor target. CDK4/6 as anti-tumor target has the following advantages: (1) for most of proliferative cells, their proliferation is CDK2 or CDK4/6-dependent, however, CDK4/6 inhibitors do not exhibit the cytotoxicity of “pan-CDK inhibitors”, such as bone marrow depression and intestinal reaction; and (2) preclinical tests show that if cyclin D level is increased or P16INK4a is inactivated in cells, the sensitivity of cells to drugs can be increased; since tumor cells have said phenomena relative to normal cells, the targeting property of drugs is increased to some extent.
So far, no CDK inhibitor drugs are approved for commercial marketing. A series of CDK4/6 inhibitors with good selectivity, which have been reported by some pharmaceutical companies including Pfizer, Eli Lilly and Novartis, are in clinical trials. Among them, of particular concern are PD0332991 (palbociclib) developed by Pfizer, LY2835219 (Phase III) developed by Eli Lilly and LEE-011 (Phase III) developed by Novartis

In April, 2013, Pfizer's PD0332991 received Breakthrough Therapy Designation from Food And Drug Administration (FDA); and in August, 2014, Pfizer submitted with FDA a New Drug Application (NDA) intended for approval of PD0332991 (palbociclib) in combination with letrozole as treatment of post-menopausal women with locally advanced or metastatic breast cancer, who are estrogen receptor-positive (ER+) and human epidermal growth factor 2 negative (HER2−), and did not received systemic treatment previously. It has very positive effect on the development of CDK4/6 inhibitors.
In order to achieve a better therapeutic effect for tumor and to better meet the market demand, the inventors hope to develop a new generation of CDK4/6 inhibitors with high efficacy and low toxicity. The invention provides selective CDK4/6 inhibitors with a new structure, and finds that the compounds with such a structure have good efficacy, and can effectively pass through the blood brain barrier, which makes CDK inhibitors as therapy for brain cancer possible.
Contents of Invention
In an aspect, the invention relates to an inhibitor/compound targeting CDK4/6 kinase. Particularly, the exemplified technical solutions of the invention are as follows:
1. A compound of Formula (I′), or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers,

wherein:
A1 and A2 each are independently selected from nitrogen;
R1 is selected from C1-6alkyl, C1-6alkoxy or 3-8 membered cycloalkyl optionally substituted by Q1, wherein Q1 is selected from C1-6alkyl or C1-6alkoxy;
R2 is selected from C1-6alkyl, C1-6alkoxy, cyano, carbamoyl or C1-6alkylcarbonylamino;
R3 and R5 each are independently selected from halogen or hydrogen, and at least one of R3 and R5 is halogen;
R4 is selected from 3-8 membered heterocyclyl, 6-14 membered fused heterocyclyl, 5-8 membered heteroaryl, 6-14 membered fused heteroaryl, phenyl, naphthyl, 6-12 membered bridged heterocyclyl or 6-12 membered spiroheterocyclyl, each of which is optionally substituted by Q2;
Q2 is selected from amino, hydroxyl, halogen, trifluoromethyl, cyano, C1-6alkoxy, C1-6alkylsulfonyl, C1-6alkylsulfonylamino, or di-C1-6alkylamino; or C1-6alkyl, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl or 6-9 membered bridged heterocyclyl, each optionally substituted by a substituent, wherein the substituent is selected from amino, hydroxyl, halogen, trifluoromethyl, cyano, C1-6alkyl, C1-6alkoxy, C1-6alkylamino, di-C1-6alkylamino, C1-6alkylsulfonyl, 3-8 membered heterocyclyl or 3-8 membered cycloalkyl;
n is selected from 0, 1, 2, 3, 4 or 5.
2. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 1, wherein
A1 and A2 each are independently selected from nitrogen;
R1 is selected from C1-4alkyl or C1-4alkoxy;
R2 is selected from C1-4alkyl, C1-4alkoxy, cyano, carbamoyl or C1-4alkylcarbonylamino;
R3 and R5 each are independently selected from halogen;
R4 is selected from a nitrogen-containing 5-6 membered heterocyclyl optionally substituted by Q2; wherein the “nitrogen-containing 5-6 membered heterocyclyl” is preferably “a nitrogen-containing 6 membered heterocyclyl”;
Q2 is selected from amino, hydroxyl, halogen, trifluoromethyl, cyano, C1-4alkoxy, or di-C1-4alkylamino; or C1-4alkyl, 3-6 membered cycloalkyl or 3-6 membered heterocyclyl, each optionally substituted by a substituent, wherein the substituent is selected from amino, hydroxyl, halogen, trifluoromethyl, C1-4alkyl, C1-4alkoxy, C1-4alkylamino, di-C1-4alkylamino, or 3-6 membered cycloalkyl;
n is selected from 0.
3. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 2, wherein the compound is selected from:

4. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 1, wherein the compound has the structure of Formula (I),

wherein:
A1 and A2 each are independently selected from nitrogen;
R1 is selected from C1-6alkyl, C1-6alkoxy or 3-8 membered cycloalkyl optionally substituted by Q1, wherein Q1 is selected from C1-6alkyl or C1-6alkoxy;
R2 is selected from C1-6alkyl, C1-6alkoxy, cyano, carbamoyl or C1-6alkylcarbonylamino;
R3 and R5 each are independently selected from halogen or hydrogen, and at least one of R3 and R5 is halogen;
R4 is selected from 3-8 membered heterocyclyl, 6-14 membered fused heterocyclyl, 5-8 membered heteroaryl, 6-14 membered fused heteroaryl, phenyl, naphthyl, 6-12 membered bridged heterocyclyl or 6-12 membered spiroheterocyclyl, each optionally substituted by Q2; wherein Q2 is selected from amino, hydroxyl, halogen, trifluoromethyl, cyano, C1-6alkyl, C1-6alkoxy, 3-8 membered heterocyclyl or 6-9 membered bridged heterocyclyl.
5. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 4, wherein
A1 and A2 each are independently selected from nitrogen;
R1 is selected from C1-4alkyl or C1-4alkoxy;
R2 is selected from C1-4alkyl, C1-4alkoxy, cyano, carbamoyl or C1-4alkylcarbonylamino;
R3 and R5 each are independently selected from halogen;
R4 is selected from 5-7 membered heterocyclyl, 6-11 membered fused heterocyclyl, 6-11 membered bridged heterocyclyl or 6-11 membered spiroheterocyclyl, each optionally substituted by Q2; wherein Q2 is selected from amino, hydroxyl, trifluoromethyl, cyano, C1-4alkyl, C1-4alkoxy, 5-6 membered heterocyclyl or 7-9 membered bridged heterocyclyl.
6. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 5, wherein
A1 and A2 each are independently selected from nitrogen;
R1 is isopropyl;
R2 is selected from methyl, methoxy, cyano, carbamoyl, or acetylamino;
R3 and R5 each are independently F;
R4 is selected from 5-6 membered heterocyclyl optionally substituted by Q2; wherein Q2 is selected from amino, hydroxyl, trifluoromethyl, cyano, C1-4alkyl, C1-4alkoxy, 6 membered heterocyclyl or 8 membered bridged heterocyclyl.
7. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 6, wherein
R2 is methyl;
R4 is selected from a nitrogen-containing 5-6 membered heterocyclyl optionally substituted by Q2; wherein the nitrogen-containing 5-6 membered heterocyclyl is linked to the methylene of Formula (I) via a nitrogen atom, wherein Q2 is selected from amino, hydroxyl, trifluoromethyl, cyano, C1-4alkyl, C1-4alkoxy, or a nitrogen-containing 8 membered bridged heterocyclyl;
wherein the nitrogen-containing 5-6 membered heterocyclyl is preferably a nitrogen-containing 5-6 membered heterocyclyl containing 1 to 2 nitrogen atoms;
8. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 7, wherein
R4 is selected from
each optionally substituted by Q2, wherein Q2 is selected from C1-4alkyl or a nitrogen-containing 8 membered bridged heterocyclyl.
9. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 5, wherein
A1 and A2 each are independently selected from nitrogen;
R1 is isopropyl;
R2 is selected from methyl, methoxy, cyano, carbamoyl, or acetylamino;
R3 and R5 each are independently F;
R4 is selected from 7-9 membered bridged heterocyclyl optionally substituted by Q2; wherein Q2 is selected from amino, hydroxyl, trifluoromethyl, cyano, C1-4alkyl, 6 membered heterocyclyl or 8 membered bridged heterocyclyl.
10. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 9, wherein
R2 is methyl;
R4 is selected from a nitrogen-containing 7-9 membered bridged heterocyclyl optionally substituted by Q2; wherein the nitrogen-containing 7-9 membered bridged heterocyclyl is linked to the methylene of Formula (I) via a nitrogen atom, wherein Q2 is selected from amino, hydroxyl, trifluoromethyl, cyano, C1-4alkyl, or a nitrogen-containing 6 membered heterocyclyl;
wherein the nitrogen-containing 7-9 membered bridged heterocyclyl is preferably a nitrogen-containing 7-9 membered bridged heterocyclyl containing 1 to 2 nitrogen atoms.
11. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 10, wherein
R4 is selected from
each optionally substituted by Q2, wherein Q2 is selected from C1-4alkyl or a nitrogen-containing 6 membered heterocyclyl.
12. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 5, wherein
A1 and A2 each are independently selected from nitrogen;
R1 is isopropyl;
R2 is selected from methyl, methoxy, cyano, carbamoyl, or acetylamino;
R3 and R5 each are F;
R4 is selected from 6-10 membered fused heterocyclyl optionally substituted by Q2; wherein Q2 is selected from amino, hydroxyl, trifluoromethyl, cyano, C1-4alkyl, 6 membered heterocyclyl or 8 membered bridged heterocyclyl.
13. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 12, wherein
R2 is methyl;
R4 is selected from a nitrogen-containing 6-10 membered fused heterocyclyl that contains 1, 2 or 3 identical or different heteroatoms and is optionally substituted by Q2; wherein the heteroatoms are preferably selected from nitrogen atom and oxygen atom, and contain at least one nitrogen atom, and the 6-10 membered fused heterocyclyl is linked to the methylene of Formula (I) via a nitrogen atom, wherein Q2 is selected from amino, hydroxyl, trifluoromethyl, cyano, or C1-4alkyl.
14. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 13 wherein
R4 is selected from
each optionally substituted by Q2, wherein Q2 is selected from amino or C1-4alkyl.
15. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 5, wherein
A1 and A2 each are independently selected from nitrogen;
R1 is isopropyl;
R2 is selected from methyl, methoxy, cyano, carbamoyl, or acetylamino;
R3 and R5 each are F;
R4 is selected from 7-11 membered spiroheterocyclyl optionally substituted by Q2; wherein Q2 is selected from amino, hydroxyl, trifluoromethyl, cyano, C1-4alkyl, 6 membered heterocyclyl or 8 membered bridged heterocyclyl.
16. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 15, wherein
R2 is selected from methyl;
R4 is selected from a nitrogen-containing 7-11 membered spiroheterocyclyl optionally substituted by Q2; wherein the nitrogen-containing 7-11 membered spiroheterocyclyl is linked to the methylene of Formula (I) via a nitrogen atom, wherein Q2 is selected from amino, hydroxyl, trifluoromethyl, cyano, of C1-4alkyl;
wherein the nitrogen-containing 7-11 membered spiroheterocyclyl is preferably a nitrogen-containing 7-11 membered spiroheterocyclyl containing 1 to 2 nitrogen atoms.
17. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 16, wherein
R4 is selected from
each optionally substituted by Q2, wherein Q2 is selected from C1-4alkyl.
18. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 1, wherein the compound is selected from the compounds shown in Table A.
TABLE AA part of compounds of the inventionNo.Formula 1  2  3  4  5  6  6-1  6-2  6-2-1  7  7-1  8  8-1  9  9-1 10 11 12 13 14 15 16 17 18 19 20 21 22 23 23-1 24 24-1 25 25-1 26 26-1 27 27-1 28 29 30 30-1 31 31-1 32 32-1 33 33-1 34 35
The invention also relates to uses of the disclosed compounds. Therefore, the invention also relates to the following exemplified technical solutions:
19. A pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to any one of Solutions 1-18, and optionally one or more pharmaceutically acceptable carriers.
20. The pharmaceutical composition according to Solution 19, further comprising one or more additional anti-tumor agents and/or immunosuppressors.
21. The pharmaceutical composition according to Solution 20, wherein the additional anti-tumor agents and/or immunosuppressors are selected from one or more of: methotrexate, capecitabine, gemcitabine, doxifluridine, pemetrexed disodium, pazopanib, imatinib, erlotinib, lapatinib, gefitinib, vandetanib, herceptin, bevacizumab, rituximab, trastuzumab, paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxycamptothecine, mitomycin, epirubicin, pirarubicin, bleomycin, letrozole, tamoxifen, fulvestrant, triptorelin, flutamide, leuprorelin, anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, mechlorethamine, melphalan, chlorambucil, carboplatin, cisplatin, oxaliplatin, lobaplatin, topotecan, camptothecin, topotecan, everolimus, sirolimus, temsirolimus, 6-mercaptopurine, 6-thioguanine, azathioprine, Actinomycin D, daunorubicin, adriamycin, mitoxantrone, bleomycin, mithramycin and aminoglutethimide.
22. Use of the compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to any one of Solutions 1-18, in the manufacture of a medicament for treating and/or preventing a cancer-related disease mediated by CDK4/6 kinase in a subject.
23. The use according to Solution 22, wherein the cancer-related disease is selected from brain tumor, lung cancer, squamous carcinoma, bladder carcinoma, gastric cancer, ovarian cancer, peritoneal carcinoma, pancreatic carcinoma, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, liver cancer, renal carcinoma, esophageal adenocarcinoma, esophageal squamous cancer, prostatic cancer, female reproductive duct cancer, cancer in situ, lymphoma, neurofibromas, thyroid carcinoma, osteocarcinoma, skin cancer, brain cancer, colon cancer, testiculus cancer, gastrointestinal stromal tumor, prostate neoplasms, mast cell tumor, multiple myeloma, melanoma, glioma or sarcoma.
24. The use according to Solution 22 or 23, wherein the subject is mammal, such as bovine, equine, caprid, suidae, canine, feline, rodent, and primate; wherein the particularly preferred subject is human.
25. The use according to any one of Solutions 22-24, wherein the medicament further comprises one or more additional anti-tumor agents and/or immunosuppressors; preferably, the additional anti-tumor agents and/or immunosuppressors are selected from one or more of: methotrexate, capecitabine, gemcitabine, doxifluridine, pemetrexed disodium, pazopanib, imatinib, erlotinib, lapatinib, gefitinib, vandetanib, herceptin, bevacizumab, rituximab, trastuzumab, paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxycamptothecine, mitomycin, epirubicin, pirarubicin, bleomycin, letrozole, tamoxifen, fulvestrant, triptorelin, flutamide, leuprorelin, anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, mechlorethamine, melphalan, chlorambucil, carboplatin, cisplatin, oxaliplatin, lobaplatin, topotecan, camptothecin, topotecan, everolimus, sirolimus, temsirolimus, 6-mercaptopurine, 6-thioguanine, azathioprine, Actinomycin D, daunorubicin, adriamycin, mitoxantrone, bleomycin, mithramycin and aminoglutethimide.
26. A method for treating and/or preventing a cancer-related disease mediated by CDK4/6 kinase, comprising administering to a subject in need thereof a therapeutically and/or prophylactically effective amount of the compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to any one of Solutions 1-18 or the pharmaceutical composition according to any one of Solutions 19-21.
27. The method according to Solution 26, wherein the cancer-related disease is selected from brain tumor, lung cancer, squamous carcinoma, bladder carcinoma, gastric cancer, ovarian cancer, peritoneal carcinoma, pancreatic carcinoma, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, liver cancer, renal carcinoma, esophageal adenocarcinoma, esophageal squamous cancer, prostatic cancer, female reproductive duct cancer, cancer in situ, lymphoma, neurofibromas, thyroid carcinoma, osteocarcinoma, skin cancer, brain cancer, colon cancer, testiculus cancer, gastrointestinal stromal tumor, prostate neoplasms, mast cell tumor, multiple myeloma, melanoma, glioma or sarcoma.
28. The method according to Solution 26 or 27, wherein the subject is mammal, such as bovine, equine, caprid, suidae, canine, feline, rodent, and primate; wherein the particularly preferred subject is human.
29. The method according to any one of Solutions 25-28, wherein the method further comprises administering to the subject one or more additional anti-tumor agents and/or immunosuppressors; preferably the additional anti-tumor agents and/or immunosuppressors are selected from one or more of: methotrexate, capecitabine, gemcitabine, doxifluridine, pemetrexed disodium, pazopanib, imatinib, erlotinib, lapatinib, gefitinib, vandetanib, herceptin, bevacizumab, rituximab, trastuzumab, paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxycamptothecine, mitomycin, epirubicin, pirarubicin, bleomycin, letrozole, tamoxifen, fulvestrant, triptorelin, flutamide, leuprorelin, anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, mechlorethamine, melphalan, chlorambucil, carboplatin, cisplatin, oxaliplatin, lobaplatin, topotecan, camptothecin, topotecan, everolimus, sirolimus, temsirolimus, 6-mercaptopurine, 6-thioguanine, azathioprine, Actinomycin D, daunorubicin, adriamycin, mitoxantrone, bleomycin, mithramycin and aminoglutethimide.
30. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to any one of Solutions 1-18, for use in the treatment and/or prevention of a cancer-related disease mediated by CDK4/6 kinase in a subject.
31. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 30, wherein the cancer-related disease is selected from brain tumor, lung cancer, squamous carcinoma, bladder carcinoma, gastric cancer, ovarian cancer, peritoneal carcinoma, pancreatic carcinoma, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, liver cancer, renal carcinoma, esophageal adenocarcinoma, esophageal squamous cancer, prostatic cancer, female reproductive duct cancer, cancer in situ, lymphoma, neurofibromas, thyroid carcinoma, osteocarcinoma, skin cancer, brain cancer, colon cancer, testiculus cancer, gastrointestinal stromal tumor, prostate neoplasms, mast cell tumor, multiple myeloma, melanoma, glioma or sarcoma.
32. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 30 or 31, wherein the subject is mammal, such as bovine, equine, caprid, suidae, canine, feline, rodent, and primate; wherein the particularly preferred subject is human.
33. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to any one of Solutions 30-32, which is used in combination with one or more additional anti-tumor agents and/or immunosuppressors; preferably the additional anti-tumor agents and/or immunosuppressors are selected from one or more of: methotrexate, capecitabine, gemcitabine, doxifluridine, pemetrexed disodium, pazopanib, imatinib, erlotinib, lapatinib, gefitinib, vandetanib, herceptin, bevacizumab, rituximab, trastuzumab, paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxycamptothecine, mitomycin, epirubicin, pirarubicin, bleomycin, letrozole, tamoxifen, fulvestrant, triptorelin, flutamide, leuprorelin, anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, mechlorethamine, melphalan, chlorambucil, carboplatin, cisplatin, oxaliplatin, lobaplatin, topotecan, camptothecin, topotecan, everolimus, sirolimus, temsirolimus, 6-mercaptopurine, 6-thioguanine, azathioprine, Actinomycin D, daunorubicin, adriamycin, mitoxantrone, bleomycin, mithramycin and aminoglutethimide.
34. Use of the compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to any one of Solutions 1-18, in the manufacture of a formulation for reducing and/or inhibiting CDK4 and/or CDK6 kinase activity in a cell.
35. The use according to Solution 34, wherein the formulation is administered in vivo or in vitro; for example, the formulation is administered to a subject (e.g., mammal, such as bovine, equine, caprid, suidae, canine, feline, rodent, and primate; e.g., human), to reduce or inhibit CDK4 and/or CDK6 kinase activity in a cell of the subject; or the formulation is administered to an in vitro cell (e.g., a cell line or a cell from a subject such as a cancer cell), to reduce or inhibit CDK4 and/or CDK6 kinase activity in the in vitro cell.
36. The use of Solution 34 or 35, wherein the cell is selected from brain tumor cells, lung cancer cells, squamous carcinoma cells, bladder carcinoma cells, gastric cancer cells, ovarian cancer cells, peritoneal carcinoma cells, pancreatic carcinoma cells, breast cancer cells, head and neck cancer cells, cervical cancer cells, endometrial cancer cells, rectal cancer cells, liver cancer cells, renal carcinoma cells, esophageal adenocarcinoma cells, esophageal squamous cancer cells, prostatic cancer cells, female reproductive duct cancer cells, cancer in situ cells, lymphoma cells, neurofibromas cells, thyroid carcinoma cells, osteocarcinoma cells, skin cancer cells, brain cancer cells, colon cancer cells, testiculus cancer cells, gastrointestinal stromal tumor cells, prostate neoplasms cells, mast cell tumor cells, multiple myeloma cells, melanoma cells, glioma cells or sarcoma cells.
37. A method for reducing or inhibiting CDK4 and/or CDK6 kinase activity in a cell, comprising administering to the cell an effective amount of the compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to any one of Solutions 1-18.
38. The method according to Solution 37, wherein the method is performed in vivo, for example, the cell is a cell in a subject (e.g., mammal, such as bovine, equine, caprid, suidae, canine, feline, rodent, and primate; e.g., human); or the method is performed in vitro, for example, the cell is an in vitro cell (e.g., a cell line or a cell from a subject such as a cancer cell).
39. The method of Solution 37 or 38, wherein the cell is selected from brain tumor cells, lung cancer cells, squamous carcinoma cells, bladder carcinoma cells, gastric cancer cells, ovarian cancer cells, peritoneal carcinoma cells, pancreatic carcinoma cells, breast cancer cells, head and neck cancer cells, cervical cancer cells, endometrial cancer cells, rectal cancer cells, liver cancer cells, renal carcinoma cells, esophageal adenocarcinoma cells, esophageal squamous cancer cells, prostatic cancer cells, female reproductive duct cancer cells, cancer in situ cells, lymphoma cells, neurofibromas cells, thyroid carcinoma cells, osteocarcinoma cells, skin cancer cells, brain cancer cells, colon cancer cells, testiculus cancer cells, gastrointestinal stromal tumor cells, prostate neoplasms cells, mast cell tumor cells, multiple myeloma cells, melanoma cells, glioma cells or sarcoma cells.
40. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to any one of Solution 1-18, for use in reduction or inhibition of CDK4 and/or CDK6 kinase activity in a cell.
41. The compound, or a pharmaceutically acceptable salt, ester, solvate or isomer thereof according to Solution 40, which is used for in vivo or in vitro administration; for example, the formulation is administered to a subject (e.g., mammal, such as bovine, equine, caprid, suidae, canine, feline, rodent, and primate; e.g., human), to reduce or inhibit CDK4 and/or CDK6 kinase activity in a cell of the subject; or the formulation is administered to an in vitro cell (e.g., a cell line or a cell from a subject such as a cancer cell), to reduce or inhibit CDK4 and/or CDK6 kinase activity in the in vitro cell.
42. The compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to Solution 40 or 41, wherein the cell is selected from brain tumor cells, lung cancer cells, squamous carcinoma cells, bladder carcinoma cells, gastric cancer cells, ovarian cancer cells, peritoneal carcinoma cells, pancreatic carcinoma cells, breast cancer cells, head and neck cancer cells, cervical cancer cells, endometrial cancer cells, rectal cancer cells, liver cancer cells, renal carcinoma cells, esophageal adenocarcinoma cells, esophageal squamous cancer cells, prostatic cancer cells, female reproductive duct cancer cells, cancer in situ cells, lymphoma cells, neurofibromas cells, thyroid carcinoma cells, osteocarcinoma cells, skin cancer cells, brain cancer cells, colon cancer cells, testiculus cancer cells, gastrointestinal stromal tumor cells, prostate neoplasms cells, mast cell tumor cells, multiple myeloma cells, melanoma cells, glioma cells or sarcoma cells.
43. A kit for reducing or inhibiting CDK4 and/or CDK6 kinase activity in a cell, comprising the compound, or a pharmaceutically acceptable salt, ester, or solvate thereof, or their stereoisomers according to any one of Solutions 1-18, and optionally an instruction.